应用

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用

Ab评论

说明

Flow Cyt

1/50.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ELISA

1/10000.

IHC-P

1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Recommended antigen retrieval:
Place slides in a special glass slide holder and fill in the rack with slides to ensure even heating.
Rinse rack in 1L of Citrate buffer in ordinary pressure cooker,(0.01M,pH6.0) to immerse all slides.
Cover the cap and heat for 15-20 minutes on electric cooker until water boils.
Begin timing, only 2 minutes in high-pressure
Remove the pressure cooker, Allow it to cool(Cool the slides at room temperature)
Rinse the slides in PBS for 5 minutes

ab54210 staining CDKN2A/p16INK4a in Human cervix and ovarian carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/4000 in Dako diluent) for 20 minutes at 25°C. An undiluted HRP-conjugated Mouse polymer was used as the secondary antibody.

ab54210 staining CDKN2A/p16INK4a in Human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 1 hour at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in 1% NGS/TBS) for 20 hours at 4°C. Ab6785 (1/800) was used as the secondary antibody.

Overlay histogram showing HeLa cells stained with ab54210 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54210, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

Here are the links to the Human Protein Atlas, including ones to the sites for normal and cancer tissue as well as cell lines. Several brain cell lines are listed as expressing CDKN2A in moderate levels.

Thank you for your enquiry and your interest in our products. We have several antibodies against CDKN2A but none of them have been tested for cross-reactivity with Syrian hamster (Mesocricetus auratus). Regarding ab54210 - the exact immunogen...